Non-invasive skin collection system

ABSTRACT

The subject matter described herein provides non-invasive tape stripping methods for the collection of a skin sample. The tape stripping method includes applying and removing at least one adhesive tape, provided that a skin sample is adhered to the adhesive tape after removal. The at least one adhesive tape is supplied in an adhesive skin sample collection kit. The adhesive skin sample collection kit further comprises a sample collector and instructions for use sheet.

CROSS-REFERENCE

This application is a continuation of U.S. application Ser. No.17/002,676 filed Aug. 25, 2020, which is a continuation of U.S.application Ser. No. 16/886,611 filed May 28, 2020, which is acontinuation of U.S. application Ser. No. 15/571,247, filed Nov. 1,2017, now issued as U.S. Pat. No. 10,709,428 on Jul. 14, 2020, which isa U.S. National Phase of International Application No.PCT/US2016/030287, filed Apr. 29, 2016, which claims the benefit of U.S.Provisional Application No. 62/156,091, filed May 1, 2015, each of whichis incorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

Skin diseases are some of the most common human illnesses and representan important global burden in healthcare. Three skin diseases are in thetop ten most prevalent diseases worldwide, and eight fall into the top50. When considered collectively, skin conditions range from being thesecond to the 11th leading causes of years lived with disability.

SUMMARY OF THE INVENTION

Skin diseases include eczema, psoriasis, acne vulgaris, pruritus,alopecia areata, decubitus ulcer, urticaria, scabies, fungal skindiseases, impetigo, abscess, bacterial skin diseases, cellulitis, lupus,viral warts, molluscum contagiosum and cancers, such as melanomas.Indeed, the deadliest skin cancer is melanoma. Melanoma is currently thefastest growing cancer with the incidence rate of melanoma havingdoubled since 1973. While there has been a 20% decline in cancer deathsoverall since 1991, melanoma is one of three cancers facing increasingrates of death. Because approximately 62% of melanomas and 45% ofmelanoma deaths occur prior to age 65, melanoma places significantburdens on the healthcare system. If diagnosed and removed early in itsevolution, when confined to the outermost skin layer and deemed to benon-invasive or “in situ” (Stage 0), patients have an expected survivalrate of almost 100%. Invasive melanomas that are thin and extend intothe uppermost regions of the second skin layer still have cure ratesgreater than 90%. However, once the cancer advances into the deeperlayers of skin, the risk of metastasis increases.

The inventors of the present disclosure have identified a need forimprovement of skin disease prevention, diagnosis and treatment toimprove patient well-being and alleviate burdens on global healthefforts. As described herein, pigmented skin lesions suspicious formelanoma can be assessed for diagnosis by both visual observation andbiopsy. Current tools commonly used to aid the biopsy decision havemodest sensitivity and low specificity and include clinical gradingcriteria (eg., ABCDE attributes) and lesion magnification with adermatoscope. Visible observation and pathologic assessment of theselesions is challenging, making melanoma one of the top five misdiagnosedcancers. Similar to visual evaluation of pigmented lesions,histopathologic assessment of pigmented lesions is also subjective andchallenging. Because patients can have many atypical lesions, thedecision of which lesions to biopsy can be challenging and it is notpractical to biopsy all lesions. Pigmented lesions with a lowersuspicion for melanoma, and likely earlier stages, may not be selectedfor biopsy such that 10%-30% may have a delayed diagnosis. Of themillions of surgical biopsies performed each year on selected pigmentedskin lesions, over 95% are negative for melanoma and represent anunnecessary surgical procedure.

Lesions positive for melanoma are subsequently staged to determine ifthe tumor remains in situ or if it has undergone invasion. For earlymelanoma, this staging can be difficult and is also dependent on thearea selected for examination. It is important to identify the stageaccurately, because invasive melanoma has a lower survival rate,requires more extensive medical treatment, surveillance, work up and hasa higher cost. The medical work up for invasive melanoma may include asentinel lymph node biopsy surgical procedure to determine if themelanoma has metastasized. This is a significant surgical procedure withassociated high cost and morbidity with results that can be ambiguous.

Provided herein are methods and systems for non-invasive skin samplingusing an adhesive tape. According to one feature of the subject matterdescribed herein, an adhesive tape having a first central collectionarea and a second area extending from the periphery of the firstcollection area is provided. The first central collection area of theadhesive tapes has a skin facing surface comprising an adhesive matrix.In some embodiments, the first central collection area of the adhesivetape has a second surface opposite the surface comprising the adhesivematrix. The second area of the adhesive tape is useful as a tab, toapply and remove the adhesive tape from a skin surface. The adhesivetape is configured for application to a skin surface so that aneffective amount of a skin sample adheres to the adhesive matrix. Insome embodiments, the effective amount of the skin sample comprises nomore than about 1 microgram of cellular material. In some embodiments,the effective amount of the skin sample comprises from about 50microgram to about 1 gram of cellular material. In some embodiments, theeffective amount of the skin sample comprises between about 50 microgramto about 500 microgram, between about 100 microgram to about 450microgram, between about 100 microgram to about 350 microgram, betweenabout 100 microgram to about 300 microgram, between about 120 microgramto about 250 microgram, or between about 150 microgram to about 200microgram of cellular material. In some embodiments, an effective amountof a skin sample is an amount sufficient to isolate and identify thecellular material. In some embodiments, the adhered skin samplecomprises a cellular material that removably adheres to the adhesivetape. In some embodiments, the cellular material is a nucleic acid suchas RNA or DNA. In some embodiments, the effective amount of the skinsample comprises between about 50 microgram to about 500 microgram,between about 100 microgram to about 450 microgram, between about 100microgram to about 350 microgram, between about 100 microgram to about300 microgram, between about 120 microgram to about 250 microgram, orbetween about 150 microgram to about 200 microgram of RNA material. Insome embodiments, the cellular material is no more than about 1 nanogramof RNA material.

In some embodiments, the adhesive tape does not contain latex, silicone,or does not contain either of these agents. In some embodiments, thematrix of the adhesive tape is comprised of a synthetic rubber compound.In some embodiments, the first collection area of the adhesive tape iscomprised of a transparent material. In some embodiments, the secondarea of the adhesive tape is comprised of a transparent material. Insome embodiments, the adhesive tape is comprised of a flexible material.In some embodiments, the first central collection area and the secondarea are comprised of different materials. In some embodiments, thefirst central collection area of the adhesive tape is comprised of apolyurethane carrier film. In some embodiments, the first centralcollection area of the adhesive tape has an elliptical shape. In someembodiments, the longest length of the first central collection area isfrom about 5 mm to about 50 mm.

According to one feature of the subject matter described herein, anadhesive tape is provided on a tri-fold skin sample collector configuredto hold the adhesive tape. The adhesive tape has a first centralcollection area and a second area extending from the periphery of thefirst collection area is provided. The first central collection area ofthe adhesive tapes has a skin facing surface comprising an adhesivematrix. In some embodiments, the first central collection area of theadhesive tape has a second surface opposite the surface comprising theadhesive matrix. The second area of the adhesive tape is useful as atab, for applying and removing the adhesive tape from a skin surface.The adhesive tape is configured for application to a skin surface sothat an effective amount of a skin sample adheres to the adhesivematrix. In some embodiments, the effective amount of the skin samplecomprises no more than about 1 microgram of cellular material. In someembodiments, an effective amount of a skin sample is an amountsufficient to isolate and identify the cellular material. In someembodiments, the effective amount of the skin sample comprises fromabout 50 microgram to about 1 gram of cellular material. In someembodiments, the effective amount of the skin sample comprises betweenabout 50 microgram to about 500 microgram, between about 100 microgramto about 450 microgram, between about 100 microgram to about 350microgram, between about 100 microgram to about 300 microgram, betweenabout 120 microgram to about 250 microgram, or between about 150microgram to about 200 microgram of cellular material. In someembodiments, the adhered skin sample comprises a cellular material thatremovably adheres to the adhesive tape.

In some embodiments, the tri-fold skin sample collector comprises threepanels. The tri-fold skin sample collector includes a peelable releasepanel and a placement area panel. In some embodiments, one panel of thetri-fold skin sample collector is a clear panel. In some embodiments,the tri-fold skin sample collector is labeled with a unique barcode thatis assigned to a patient sample. In some embodiments, the placement areapanel of the tri-fold skin sample collector comprises a removable liner.In some embodiments, the adhesive tape is affixed to the peelablerelease panel prior to skin application. In some embodiments, thepeelable release panel of the tri-fold skin sample collector isconfigured to hold between about 1 to about 12 adhesive tapes, betweenabout 2 to about 12 adhesive tapes, 1 to about 8 adhesive tapes, between4 to 10 adhesive tapes, between 6 to 10 adhesive tapes, between 6 to 8adhesive tapes, or between 4 to 8 adhesive tapes provided that theadhesive tapes have not been applied to a skin surface. In someembodiments, the peelable release panel is configured to hold 8 adhesivetapes, provided that the adhesive tapes have not been applied to a skinsurface. In some embodiments, the peelable release panel is configuredto hold 4 adhesive tapes, provided that the adhesive tapes have not beenapplied to a skin surface. In some embodiments, the adhesive tape isaffixed to the placement panel after skin application, provided that theadhesive tape comprises an effective amount of a skin sample. In someembodiments, the placement area panel of the tri-fold skin samplecollector is configured to hold between about 1 to about 12 adhesivetapes, between about 2 to about 12 adhesive tapes, 1 to about 8 adhesivetapes, between 4 to 10 adhesive tapes, between 6 to 10 adhesive tapes,between 6 to 8 adhesive tapes, or between 4 to 8 adhesive tapes providedthat each adhesive tape comprises an effective amount of a skin sample.In some embodiments, the placement area panel of the tri-fold skinsample collector is configured to hold 8 adhesive tapes, provided thateach adhesive tape comprises an effective amount of a skin sample. Insome embodiments, the placement area panel of the tri-fold skin samplecollector is configured to hold 4 adhesive tapes, provided that eachadhesive tape comprises an effective amount of a skin sample.

According to one aspect of the subject matter, a non-invasive method forisolating a skin sample is provided. The skin sample is isolated byapplying an adhesive tape to a desired skin surface, where the skinsample adheres to the adhesive matrix, and removing the adhesive tape,stripping an adhered skin sample from the skin surface. In someembodiments, the adhesive tape is slowly removed from the skin in onedirection. In some embodiments, the person applying the adhesive tapewears gloves. The adhesive tape has a first central collection area anda second area extending from the periphery of the first collection areais provided. The first central collection area of the adhesive tapes hasa skin facing surface comprising an adhesive matrix. In someembodiments, the first central collection area of the adhesive tape hasa second surface opposite the surface comprising the adhesive matrix.The second area of the adhesive tape is useful as a tab, for applyingand removing the adhesive tape from a skin surface. The adhesive tape isconfigured for application to a skin surface so that an effective amountof a skin sample adheres to the adhesive matrix. In some embodiments,the effective amount of the skin sample comprises no more than about 1microgram of cellular material. In some embodiments, an effective amountof a skin sample is an amount sufficient to isolate and identify thecellular material. In some embodiments, the adhered skin samplecomprises a cellular material that removably adheres to the adhesivetape.

In some embodiments, the adhesive matrix of the adhesive tape iscomprised of a synthetic rubber compound. In some embodiments, theadhesive tape is comprised of a transparent material. In someembodiments, the adhesive tape is comprised of a flexible material.

In some embodiments, prior to applying the adhesive tape to a desiredskin surface, the skin facing surface of the first central collectionarea does not come in contact with a skin surface other than the desiredskin surface to be sampled. In some embodiments, prior to applying theadhesive tape to a desired skin surface, the adhesive matrix of thefirst central collection area does not come in contact with a skinsurface other than the desired skin surface to be sampled.

In some embodiments, the skin surface is prepared for skin sampling byremoving any hairs on the skin surface, cleansing the surface with anantiseptic, drying the surface completely prior to application of theadhesive tape, or any combination thereof. In some embodiments, theantiseptic comprises an alcohol. In some embodiments, the alcohol isisopropyl alcohol.

In some embodiments, the method includes first removing the adhesivetape from a peelable release panel of a tri-fold skin sample collectorprior to applying the adhesive tape to the desired skin surface to besampled. In some embodiments, the tri-fold skin sample collectorincludes three panels. In some embodiments, the tri-fold skin samplecollector includes a peelable release panel. In some embodiments, thetri-fold skin sample collector includes a placement area panel. In someembodiments, the tri-fold skin sample collector includes a clear panel.In some embodiments, the tri-fold skin sample collector is labeled witha unique barcode that is assigned to a patient skin sample. In someembodiments, the method includes filling out patient information on thetri-fold skin sample collector. In some embodiments, the method includesplacing the adhesive tape and the adhered skin sample onto a placementarea panel of the tri-fold skin sample collector, provided that theadhesive matrix side of the tape is facing toward the placement areapanel. In some embodiments, the method includes removing a removableliner from the placement area panel of the tri-fold skin samplecollector prior to placing the adhesive tape onto the placement areapanel.

In some embodiments, the method comprises applying and removing betweenabout 1 to about 12 adhesive tapes, between about 2 to about 12 adhesivetapes, 1 to about 8 adhesive tapes, between 4 to 10 adhesive tapes,between 6 to 10 adhesive tapes, between 6 to 8 adhesive tapes, orbetween 4 to 8 adhesive tapes sequentially to and from the skin surface.In some embodiments, the method comprises applying and removing 8adhesive tapes sequentially to and from the skin surface. In someembodiments, the method comprises applying and removing 4 adhesive tapessequentially to and from the skin surface.

In some embodiments, the method includes holding the skin surface tautand pressing the adhesive tape firmly on the skin surface while makingcircular motions on the adhesive tape prior to removing the adhesivetape from the skin surface. In some embodiments, 1-20 circular motionsare made on the adhesive tape. In some embodiments, 15 circular motionsare made on the adhesive tape.

In some embodiments, the adhesive tape is applied onto a skin lesion ofthe skin surface. In some embodiments, the skin lesion is a pigmentedskin lesion comprising a mole, dark colored skin spot, or melanincontaining skin area. In some embodiments, the skin lesion is suspiciousfor skin diseases including, but not limited to, melanoma, lupus,rubeola, acne, hemangioma, psoriasis, eczema, candidiasis, impetigo,shingles, leprosy, Chron's disease, inflammatory dermatoses, bullousdiseases, infections, basal cell carcinoma, actinic keratoses, merkelcell carcinoma, sebaceous carcinoma, squamous cell carcinoma, anddermatofibrosarcoma protuberans. In some embodiments, the skin lesion isfrom about 5 mm to about 20 mm in diameter.

In some embodiments, the adhesive tape is applied on a skin surface thatis not located on the areas including, but limited to, palms, soles offeet, and mucous membranes. In some embodiments, the adhesive tape isapplied to a skin surface located on the areas including, but notlimited to, the face, neck, arm, chest, abdomen, back, or legs. In someembodiments, the skin surface is not ulcerated or bleeding. In someembodiments, the skin surface has not been previously biopsied.

In some embodiments, the method further comprises applying the adhesivetape to a skin lesion on the skin surface and demarcating on theadhesive tape a zone around the skin lesion on a second surface of theadhesive tape. The second surface of the adhesive tape is the surfacewhich is not the skin facing surface and does not comprise the adhesivematrix. In some embodiments, a permanent marker is used to demarcate theskin lesion zone.

In some embodiments, the method further comprises detecting the presenceof a nucleic acid molecule expressed from C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), IL-6, IL-8, IL-17A, IL-17C,IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24, IL-26, TNF-α, TNFRSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2, DEFB4A, or acombination thereof. In some embodiments, the method comprises detectingthe presence of a nucleic acid molecule expressed from Table 1 in theskin sample. In some embodiments, the method comprises detecting thepresence of a nucleic acid molecule expressed from C6orf218 or PRAME inthe skin sample. In some embodiments, the method comprises detecting thepresence of a nucleic acid molecule expressed from C6orf218 in the skinsample.

According to one aspect of the subject matter, a system for collectingand mailing a skin sample from a patient is provided. The systemincludes, but is not limited to, at least one adhesive tape, aninstructions for use sheet (or instruction manual), and a samplecollector. The adhesive tape has a first central collection area and asecond area extending from the periphery of the first collection area isprovided. The first central collection area of the adhesive tapes has askin facing surface comprising an adhesive matrix. In some embodiments,the first central collection area of the adhesive tape has a secondsurface opposite the surface comprising the adhesive matrix. The secondarea of the adhesive tape is useful as a tab, for applying and removingthe adhesive tape from a skin surface. The adhesive tape is configuredfor application to a skin surface so that an effective amount of a skinsample adheres to the adhesive matrix. In some embodiments, theeffective amount of the skin sample comprises no more than about 1microgram of cellular material. In some embodiments, the effectiveamount of the skin sample comprises between about 50 microgram to about500 microgram, between about 100 microgram to about 450 microgram,between about 100 microgram to about 350 microgram, between about 100microgram to about 300 microgram, between about 120 microgram to about250 microgram, or between about 150 microgram to about 200 microgram ofcellular material. In some embodiments, an effective amount of a skinsample is an amount sufficient to isolate and identify the cellularmaterial. In some embodiments, the adhered skin sample comprises acellular material that removably adheres to the adhesive tape. Theinstructions for use sheet instructs for the non-invasive removal of askin sample including application of the adhesive tape to the skin, andremoval of the adhesive tape from the skin.

In some embodiments, the sample collector is a tri-fold skin samplecollector. In some embodiments, the sample collector is labeled with aunique barcode that is assigned to the patient skin sample. In someembodiments, the sample collector comprises an area for labeling patientinformation. In some embodiments, the sample collector includes apeelable release panel. In some embodiments, the sample collectorincludes a placement area panel. In some embodiments, the samplecollector includes a clear panel. In some embodiments, at least oneadhesive tape is affixed to a peelable release panel of the samplecollector. In some embodiments, between about 1 to about 12 adhesivetapes, between about 2 to about 12 adhesive tapes, 1 to about 8 adhesivetapes, between 4 to 10 adhesive tapes, between 6 to 10 adhesive tapes,between 6 to 8 adhesive tapes, or between 4 to 8 adhesive tapes adhesivetapes are affixed to a peelable release panel of the sample collector.

In some embodiments, the system includes a lab requisition form. In someembodiments, the lab requisition form is labeled with a unique barcodethat is assigned to the patient sample. In some embodiments, the systemincludes a permanent marker. In some embodiments, the system includes aresealable plastic bag. In some embodiments, the system includes apackage for shipping. In some embodiments, the package for shippingincludes a prepaid shipping label.

BRIEF DESCRIPTION OF THE DRAWINGS

The novel and inventive features of the subject matter described hereinare set forth with particularity in the appended claims. A betterunderstanding of the feature and advantages of the present inventionwill be obtained by reference to the following detailed description thatsets forth illustrative embodiments, in which the principles of theinvention are utilized, and the accompanying drawings of which:

FIG. 1 is illustrative of cleansing a skin sampling area comprising askin lesion.

FIG. 2 is illustrative of a tri-fold skin sample collector comprising apeelable release panel comprising four adhesive tapes, a placement areapanel comprising a removable liner, and a clear panel.

FIG. 3 is illustrates removing a first adhesive tape positioned at thefar left side of a peelable release panel of a tri-fold skin samplecollector.

FIG. 4 is illustrative of an adhesive tape positioned on a cleansed skinsampling area comprising a skin lesion.

FIG. 5 illustrates pressing firmly on an adhesive tape positioned on acleansed skin sampling area while making a circular motion.

FIG. 6 is illustrative of demarcating a region comprising a skin lesionon an adhesive tape.

FIG. 7 is illustrative of placing a used adhesive tape comprising a skinsample onto a placement area panel of a tri-fold skin sample collector.

FIG. 8 is illustrative of an adhesive skin sample collection kit.

DETAILED DESCRIPTION OF THE INVENTION

The subject matter described herein is based on a non-invasive tapestripping method for the collection of a skin sample. The tape strippingmethod is performed using an adhesive skin sample collection kit. Thetape stripping method involves applying and removing an adhesive tape tothe skin surface of a subject. The adhesive tape comprises an adhesivematrix, wherein during application of the adhesive tape to the skinsurface, an effective amount of a skin sample containing cellularmaterial adheres to the adhesive matrix. The adhered skin sample isretained on the adhesive matrix upon removal of the tape from the skinsurface. The adhesive tape containing the adhered skin sample isdesignated as a used adhesive tape. The adhesive tape is configured sothat at least a portion of the skin sample cellular material can beharvested from a used tape.

Non-Invasive Tape Striping and Analysis

The adhesive skin sample collection kit for use with tape strippingmethods is provided as a non-invasive means to collect skin samples withminimal discomfort. Cellular material is isolated from the skin sampleand can be utilized in tests that can determine the stage of disease,the risk of disease progression and a patient's likelihood of respondingto a particular treatment. Treatments include drug therapies and biopsy.Skin sample cellular materials include nucleic acids, polypeptides,lipids, carbohydrates and small molecules. Nucleic acids include DNA andRNA.

In some embodiments, isolated RNA from a collected skin sample isreverse transcribed into cDNA for amplification by PCR to enrich fortarget genes. The expression levels of these target genes are quantifiedby quantitative PCR in a gene expression test. A gene expression testprovides information on a gene expression signature associated with adisease. A pigmented lesion assay is an exemplary gene expression testwhich measures the expression levels of target genes from RNA isolatedusing the adhesive skin sample collection kit.

For example, in some embodiments, the pigmented lesion assay providesobjective information on a gene expression signature associated withmelanoma. This information can be used to help support a histopathologicdiagnosis or to determine the need for a biopsy, thereby reducingunnecessary biopsy procedures. The development of invasive tumorproperties is also controlled by gene expression; therefore thepigmented lesion assay may also differentiate invasive melanoma frommelanoma in situ as well as provide staging information. Theidentification of invasive melanoma with metastatic potential willdirect treatments to only those who need it. Another gene expressionassay may determine if a melanoma tumor has spread to the lymph nodes.This test can reduce the need for a sentinel lymph node surgery, whichcan be extensive, cause morbidity and has significant medical costs.

Gene expression analyses facilitate drug development by identifying drugtargets and stratifying patients into groups that will maximize a drugresponse. In an exemplary embodiment, a skin sample collected from theface of a subject with lupus is isolated and utilized in a geneexpression test to assess the expression of target genes indicated inlupus drug effects. This gene expression test can identify responders totherapy and identify new drug targets. The use of the adhesive tapeallows for skin sample collection without the scarring that can occurwith a biopsy.

In some embodiments, one or more polypeptides isolated from the usedadhesive tape are detected and/or quantified. For example, in someembodiments, one or more polypeptides isolated from the used adhesivetape are detected and/or quantified using ELISA, immunohistochemistry,mass spectrometry, and/or absorbance measurement. In some embodiments,the sequence of DNA isolated from the used adhesive tape is determinedusing gene sequencing methods known to one of skill in the art.

In some instances, the skin sample collected using the tape strippingmethod is used in combination with other clinical assays includingimmunohistochemistry, immunophenotyping, fluorescent in situhybridization (FISH), and/or any combination thereof. The skin sampledoes not necessarily need to be removed from the adhesive tape to proveuseful as an assay component. Cellular material from the skin samplescan be detected from the surface of the adhesive tape matrix. Detectionmethods include the use of probes configured to bind to cellularmaterial adhered to the adhesive tape matrix. Probes include, but arenot limited to, primers configured to bind to nucleic acids, andantibodies configured to bind to polypeptides, nucleic acids, smallmolecules, lipids, and/or carbohydrates.

In some embodiments, the tape stripping method is part of the work upfor a variety of suspected skin conditions including, but not limitedto, lupus, rubeola, acne, hemangioma, psoriasis, eczema, candidiasis,impetigo, shingles, leprosy and Chron's disease. Skin conditions alsoinclude inflammatory dermatoses, bullous diseases, infections andcancers. Skin cancers include, but are not limited to, basal cellcarcinoma, actinic keratoses, merkel cell carcinoma, sebaceouscarcinoma, squamous cell carcinoma, melanoma and dermatofibrosarcomaprotuberans.

In some embodiments, the tape stripping method is performed using aplurality of adhesive tapes. Between 1 and 8 adhesive tapes can besequentially applied and removed to collect a skin sample. The number ofadhesive tapes used per skin sample may include, but is not limited to,12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, from about 2 to about 7, fromabout 3 to about 6, and from about 4 to about 5. In certain instances,an adhesive tape is applied to the skin and removed from the skin about1 to about 8 times.

Components of the Collection Kit

The adhesive tape of the adhesive skin sample collection kit typicallycomprises a first collection area comprising an adhesive matrix and asecond area extending from the periphery of the first collection area.The adhesive matrix is located on a skin facing surface of the firstcollection area. The second area functions as a tab, suitable forapplying and removing the adhesive tape. The tab is sufficient in sizeso that while applying the adhesive tape to a skin surface, theapplicant does not come in contact with the matrix material of the firstcollection area. In some embodiments, the adhesive tape does not containa second area tab. In some instances, the adhesive tape is handled withgloves to reduce contamination of the adhesive matrix prior to use.

In some embodiments, the first collection area is a polyurethane carrierfilm. In some embodiments, the adhesive matrix is comprised of asynthetic rubber compound. In some embodiments, the adhesive matrix is astyrene-isoprene-styrene (SIS) linear block copolymer compound. In someinstances, the adhesive tape does not comprise latex, silicone, or both.In some instances, the adhesive tape is manufactured by applying anadhesive material as a liquid-solvent mixture to the first collectionarea and subsequently removing the solvent.

The matrix material is sufficiently sticky to adhere to a skin sample.The matrix material is not so sticky that is causes scarring or bleedingor is difficult to remove. In some embodiments, the matrix material iscomprised of a transparent material. In some instances, the matrixmaterial is biocompatible. In some instances, the matrix material doesnot leave residue on the surface of the skin after removal. In certaininstances, the matrix material is not a skin irritant.

In some embodiments, the adhesive tape comprises a flexible material,enabling the tape to conform to the shape of the skin surface uponapplication. In some instances, at least the first collection area isflexible. In some instances, the tab is plastic. In an illustrativeexample, the adhesive tape does not contain latex, silicone, or both. Insome embodiments, the adhesive tape is made of a transparent material,so that the skin sampling area of the subject is visible afterapplication of the adhesive tape to the skin surface. The transparencyensures that the adhesive tape is applied on the desired area of skincomprising the skin area to be sampled. In some embodiments, theadhesive tape is between about 5 and about 100 mm in length. In someembodiments, the first collection area is between about 5 and about 40mm in length. In some embodiments, the first collection area is betweenabout 10 and about 20 mm in length. In some embodiments the length ofthe first collection area is configured to accommodate the area of theskin surface to be sampled, including, but not limited to, about 19 mm,about 20 mm, about 21 mm, about 22 mm, about 23 mm, about 24 mm, about25 mm, about 30 mm, about 35 mm, about 40 mm, about 45 mm, about 50 mm,about 55 mm, about 60 mm, about 65 mm, about 70 mm, about 75 mm, about80 mm, about 85 mm, about 90 mm, and about 100 mm. In some embodiments,the first collection area is elliptical.

In further embodiments, the adhesive tape of this invention is providedon a peelable release sheet in the adhesive skin sample collection kit.In some embodiments, the adhesive tape provided on the peelable releasesheet is configured to be stable at temperatures between −80° C. and 30°C. for at least 6 months, at least 1 year, at least 2 years, at least 3years, and at least 4 years. In some instances, the peelable releasesheet is a panel of a tri-fold skin sample collector.

The peelable release sheet is configured to hold a plurality of adhesivetapes, including, but not limited to, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3,2, 1, from about 2 to about 8, from about 2 to about 7, from about 2 toabout 6, from about 2 to about 4, from about 3 to about 6, from about 3to about 8, from about 4 to about 10, from about 4 to about 8, fromabout 4 to about 6, from about 4 to about 5, from about 6 to about 10,from about 6 to about 8, or from about 4 to about 8. The peelablerelease sheet is configured to hold about 12 adhesive tapes. Thepeelable release sheet is configured to hold about 11 adhesive tapes.The peelable release sheet is configured to hold about 10 adhesivetapes. The peelable release sheet is configured to hold about 9 adhesivetapes. The peelable release sheet is configured to hold about 8 adhesivetapes. The peelable release sheet is configured to hold about 7 adhesivetapes. The peelable release sheet is configured to hold about 6 adhesivetapes. The peelable release sheet is configured to hold about 5 adhesivetapes. The peelable release sheet is configured to hold about 4 adhesivetapes. The peelable release sheet is configured to hold about 3 adhesivetapes. The peelable release sheet is configured to hold about 2 adhesivetapes. The peelable release sheet is configured to hold about 1 adhesivetape.

The adhesive tape is applied to the skin and removed from the skin.After removing the used adhesive tape from the skin surface, the tapestripping method further comprises storing the used tape on a placementarea sheet, where the tape remains until the skin sample is isolated orotherwise utilized. The used tape is configured to be stored on theplacement area sheet for at least 1 week at temperatures between −80° C.and 30° C. In some embodiments, the used tape is configured to be storedon the placement area sheet for at least 2 weeks, at least 3 weeks, atleast 1 month, at least 2 months, at least 3 months, at least 4 months,at least 5 months, and at least 6 months at temperatures between −80° C.to 30° C.

In some instances, the placement area sheet comprises a removable liner,provided that prior to storing the used tape on the placement areasheet, the removable liner is removed. The placement area sheet isconfigured to hold a plurality of adhesive tapes, including, but notlimited to, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, from about 2 to about8, from about 2 to about 7, from about 2 to about 6, from about 2 toabout 4, from about 3 to about 6, from about 3 to about 8, from about 4to about 10, from about 4 to about 8, from about 4 to about 6, fromabout 4 to about 5, from about 6 to about 10, from about 6 to about 8,or from about 4 to about 8. The placement area sheet is configured tohold about 12 adhesive tapes. The placement area sheet is configured tohold about 11 adhesive tapes. The placement area sheet is configured tohold about 10 adhesive tapes. The placement area sheet is configured tohold about 9 adhesive tapes. The placement area sheet is configured tohold about 8 adhesive tapes. The placement area sheet is configured tohold about 7 adhesive tapes. The placement area sheet is configured tohold about 6 adhesive tapes. The placement area sheet is configured tohold about 5 adhesive tapes. The placement area sheet is configured tohold about 4 adhesive tapes. The placement area sheet is configured tohold about 3 adhesive tapes. The placement area sheet is configured tohold about 2 adhesive tapes. The placement area sheet is configured tohold about 1 adhesive tape.

The used tape is stored so that the matrix containing, skin facingsurface of the used tape is in contact with the placement area sheet. Insome instances, the placement area sheet is a panel of the tri-fold skinsample collector. In some instances, the tri-fold skin sample collectormay further comprise a clear panel. The tri-fold skin sample collectormay be labeled with a unique barcode that is assigned to a subject. Insome instances, the tri-fold skin sample collector comprises an area forlabeling subject information.

In an illustrative embodiment, the adhesive skin sample collection kitcomprises the tri-fold skin sample collector comprising adhesive tapesstored on a peelable release panel. In some instances, the tri-fold skinsample collector further comprises a placement area panel with aremovable liner. The tape stripping method involves removing an adhesivetape from the tri-fold skin sample collector peelable release panel,applying the adhesive tape to a skin sample, removing the used adhesivetape containing a skin sample and placing the used tape on the placementarea sheet. In some instances the placement area panel is a singleplacement area panel sheet. The identity of the skin sample collected isindexed to the tri-fold skin sample collector or placement area panelsheet by using a barcode or printing patient information on thecollector or panel sheet. The indexed tri-fold skin sample collector orplacement sheet is sent to a diagnostic lab for processing. The usedtape is configured to be stored on the placement panel for at least 1week at temperatures between −80° C. and 25° C. In some embodiments, theused tape is configured to be stored on the placement area panel for atleast 2 weeks, at least 3 weeks, at least 1 month, at least 2 months, atleast 3 months, at least 4 months, at least 5 months, and at least 6months at temperatures between −80° C. and 25° C. In some embodimentsthe indexed tri-fold skin sample collector or placement sheet is sent toa diagnostic lab using UPS or FedEx.

In an exemplary embodiment, the tape stripping method further comprisespreparing the skin sample prior to application of the adhesive tape.Preparation of the skin sample includes, but is not limited to, removinghairs on the skin surface, cleansing the skin surface and/or drying theskin surface. In some instances, the skin surface is cleansed with anantiseptic including, but not limited to, alcohols, quaternary ammoniumcompounds, peroxides, chlorhexidine, halogenated phenol derivatives andquinolone derivatives. In some instances, the alcohol is about 0 toabout 20%, about 20 to about 40%, about 40 to about 60%, about 60 toabout 80%, or about 80 to about 100% isopropyl alcohol. In someinstances, the antiseptic is 70% isopropyl alcohol.

In some embodiments, the tape stripping method is used to collect a skinsample from the surfaces including, but not limited to, the face, head,neck, arm, chest, abdomen, back, leg, hand or foot. In some instances,the skin surface is not located on a mucous membrane. In some instances,the skin surface is not ulcerated or bleeding. In certain instances, theskin surface has not been previously biopsied. In certain instances, theskin surface is not located on the soles of the feet or palms.

The tape stripping method, devices, and systems described herein areuseful for the collection of a skin sample from a skin lesion. A skinlesion is a part of the skin that has an appearance or growth differentfrom the surrounding skin. In some instances, the skin lesion ispigmented. A pigmented lesion includes, but is not limited to, a mole,dark colored skin spot and a melanin containing skin area. In someembodiments, the skin lesion is from about 5 mm to about 16 mm indiameter. In some instances, the skin lesion is from about 5 mm to about15 mm, from about 5 mm to about 14 mm, from about 5 mm to about 13 mm,from about 5 mm to about 12 mm, from about 5 mm to about 11 mm, fromabout 5 mm to about 10 mm, from about 5 mm to about 9 mm, from about 5mm to about 8 mm, from about 5 mm to about 7 mm, from about 5 mm toabout 6 mm, from about 6 mm to about 15 mm, from about 7 mm to about 15mm, from about 8 mm to about 15 mm, from about 9 mm to about 15 mm, fromabout 10 mm to about 15 mm, from about 11 mm to about 15 mm, from about12 mm to about 15 mm, from about 13 mm to about 15 mm, from about 14 mmto about 15 mm, from about 6 to about 14 mm, from about 7 to about 13mm, from about 8 to about 12 mm and from about 9 to about 11 mm indiameter. In some embodiments, the skin lesion is from about 10 mm toabout 20 mm, from about 20 mm to about 30 mm, from about 30 mm to about40 mm, from about 40 mm to about 50 mm, from about 50 mm to about 60 mm,from about 60 mm to about 70 mm, from about 70 mm to about 80 mm, fromabout 80 mm to about 90 mm, and from about 90 mm to about 100 mm indiameter. In some instances, the diameter is the longest diameter of theskin lesion. In some instances, the diameter is the smallest diameter ofthe skin lesion.

The adhesive skin sample collection kit comprises at least one adhesivetape, a sample collector, and an instructions for use sheet. In anexemplary embodiment, the sample collector is a tri-fold skin samplecollector comprising a peelable release panel comprising at least oneadhesive tape, a placement area panel comprising a removable liner, anda clear panel. The tri-fold skin sample collector may further comprise abarcode and/or an area for transcribing patient information. Theadhesive skin sample collection kit is configured to include a pluralityof adhesive tapes, including but not limited to 12, 11, 10, 9, 8, 7, 6,5, 4, 3, 2, 1, from about 2 to about 8, from about 2 to about 7, fromabout 2 to about 6, from about 2 to about 4, from about 3 to about 6,from about 3 to about 8, from about 4 to about 10, from about 4 to about8, from about 4 to about 6, from about 4 to about 5, from about 6 toabout 10, from about 6 to about 8, or from about 4 to about 8. Theinstructions for use sheet provides the kit operator all of thenecessary information for carrying out the tape stripping method. Theinstructions for use sheet preferably includes diagrams to illustratethe tape stripping method.

In some instances, the adhesive skin sample collection kit provides allthe necessary components for performing the tape stripping method. Insome embodiments, the adhesive skin sample collection kit includes a labrequisition form for providing patient information. In some instances,the kit further comprises accessory components. Accessory componentsinclude, but are not limited to, a marker, a resealable plastic bag,gloves and a cleansing reagent. The cleansing reagent includes, but isnot limited to, an antiseptic such as isopropyl alcohol. The componentsof the skin sample collection kit may be provided in a cardboard box.

Tissue Sampling and Cellular Material

The methods, devices, and systems provided herein involve applying anadhesive or other similar tape to the skin in a manner so that aneffective or sufficient amount of a tissue, such as a skin sample,adheres to the adhesive matrix of the adhesive tape. For example, insome embodiments, the effective or sufficient amount of a skin sample isan amount that removably adheres to a material, such as the matrix oradhesive tape. The adhered skin sample, in certain embodiments,comprises cellular material including nucleic acids and proteins. Insome instances, the nucleic acid is RNA or DNA. An effective amount of askin sample contains an amount of cellular material sufficient forperforming a diagnostic assay. In some instances, the diagnostic assayis performed using the cellular material isolated from the adhered skinsample on the used adhesive tape. In some instances, the diagnosticassay is performed on the cellular material adhered to the used adhesivetape. In some embodiments, an effect amount of a skin sample comprisesan amount of RNA sufficient to perform a gene expression analysis.Sufficient amounts of RNA include picogram, nanogram, and microgramquantities.

In still further or additional embodiments, the adhered skin samplecomprises cellular material including nucleic acids such as RNA or DNA,or a polypeptide such as a protein, in an amount that is at least about1 picogram. In some embodiments, the amount of cellular material is nomore than about 1 nanogram. In further or additional embodiments, theamount of cellular material is no more than about 1 microgram. In stillfurther or additional embodiments, the amount of cellular material is nomore than about 1 gram.

In further or additional embodiments, the amount of cellular material isfrom about 1 picogram to about 1 gram. In further or additionalembodiments, the cellular material comprises an amount that is fromabout 50 microgram to about 1 gram, from about 100 picograms to about500 micrograms, from about 500 picograms to about 100 micrograms, fromabout 750 picograms to about 1 microgram, from about 1 nanogram to about750 nanograms, or from about 1 nanogram to about 500 nanograms.

In further or additional embodiments, the amount of cellular material,including nucleic acids such as RNA or DNA, or a polypeptide such as aprotein, comprises an amount that is from about 50 microgram to about500 microgram, from about 100 microgram to about 450 microgram, fromabout 100 microgram to about 350 microgram, from about 100 microgram toabout 300 microgram, from about 120 microgram to about 250 microgram,from about 150 microgram to about 200 microgram, from about 500nanograms to about 5 nanograms, or from about 400 nanograms to about 10nanograms, or from about 200 nanograms to about 15 nanograms, or fromabout 100 nanograms to about 20 nanograms, or from about 50 nanograms toabout 10 nanograms, or from about 50 nanograms to about 25 nanograms.

In further or additional embodiments, the amount of cellular material,including nucleic acids such as RNA or DNA, or a polypeptide such as aprotein, is less than about 1 gram, is less than about 500 micrograms,is less than about 490 micrograms, is less than about 480 micrograms, isless than about 470 micrograms, is less than about 460 micrograms, isless than about 450 micrograms, is less than about 440 micrograms, isless than about 430 micrograms, is less than about 420 micrograms, isless than about 410 micrograms, is less than about 400 micrograms, isless than about 390 micrograms, is less than about 380 micrograms, isless than about 370 micrograms, is less than about 360 micrograms, isless than about 350 micrograms, is less than about 340 micrograms, isless than about 330 micrograms, is less than about 320 micrograms, isless than about 310 micrograms, is less than about 300 micrograms, isless than about 290 micrograms, is less than about 280 micrograms, isless than about 270 micrograms, is less than about 260 micrograms, isless than about 250 micrograms, is less than about 240 micrograms, isless than about 230 micrograms, is less than about 220 micrograms, isless than about 210 micrograms, is less than about 200 micrograms, isless than about 190 micrograms, is less than about 180 micrograms, isless than about 170 micrograms, is less than about 160 micrograms, isless than about 150 micrograms, is less than about 140 micrograms, isless than about 130 micrograms, is less than about 120 micrograms, isless than about 110 micrograms, is less than about 100 micrograms, isless than about 90 micrograms, is less than about 80 micrograms, is lessthan about 70 micrograms, is less than about 60 micrograms, is less thanabout 50 micrograms, is less than about 20 micrograms, is less thanabout 10 micrograms, is less than about 5 micrograms, is less than about1 microgram, is less than about 750 nanograms, is less than about 500nanograms, is less than about 250 nanograms, is less than about 150nanograms, is less than about 100 nanograms, is less than about 50nanograms, is less than about 25 nanograms, is less than about 15nanograms, is less than about 1 nanogram, is less than about 750picograms, is less than about 500 picograms, is less than about 250picograms, is less than about 100 picograms, is less than about 50picograms, is less than about 25 picograms, is less than about 15picograms, or is less than about 1 picogram.

Analysis of Cellular Material and Communication of Results

In some embodiments, isolated RNA from a collected skin sample isreverse transcribed into cDNA, for example for amplification by PCR toenrich for target genes. The expression levels of these target genes arequantified by quantitative PCR in a gene expression test. In someinstances, in combination with quantitative PCR, a software programperformed on a computer is utilized to quantify RNA isolated from thecollected skin sample. In some instances, a software program or moduleis utilized to relate a quantity of RNA from a skin sample to a geneexpression signature, wherein the gene expression signature isassociated with a disease such as melanoma. In some embodiments, asoftware program or module scores a sample based on gene expressionlevels. In some embodiments, the sample score is compared with areference sample score to determine if there is a statisticalsignificance between the gene expression signature and a disease.

In some embodiments, one or more target genes from the isolated RNAobtained from a collected skin sample are analyzed. In some instances,from about 1 to about 100, from about 1 to about 90, from about 1 toabout 80, from about 1 to about 70, from about 1 to about 60, from about1 to about 50, from about 1 to about 40, from about 1 to about 30, fromabout 1 to about 20, from about 5 to about 100, from about 5 to about80, from about 5 to about 60, from about 5 to about 40, from about 5 toabout 20, from about 10 to about 100, from about 10 to about 80, fromabout 10 to about 60, from about 10 to about 40, from about 20 to about80, from about 20 to about 60, from about 20 to about 40, from about 30to about 80, from about 30 to about 60, from about 40 to about 60, fromabout 2 to about 10, from about 2 to about 8, or from about 2 to about 6target genes from the isolated RNA obtained from a collected skin sampleare analyzed.

In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,20, 25, 30, 40, 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 1 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 2 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 3 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 4 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 5 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 6 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 7 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 8 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 9 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 10 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 11 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 12 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 13 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 14 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 15 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 20 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 25 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed. Insome cases, about 30 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed. In some cases, about 40 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed. In some cases, about 50 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed.

In some embodiments, the one or more target genes comprise C6orf218,preferentially expressed antigen in melanoma (PRAME), IL-6, IL-8,IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24,IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2,DEFB4A, or a combination thereof. In some embodiments, the one or moretarget genes comprise a target gene selected from Table 1.

TABLE 1 Gene Symbol Species Gene Name IL-23A Human interleukin 23, alphasubunit p19 IL-17A Human interleukin 17 A IL-17C Human interieukin 17 CIL-17F Human interieukin 17 F TNF-α Human tumor necrosis factor IL-17RAHuman interleukin 17 receptor A IL-17RC Human interleukin 17 receptor CTNF Human tumor necrosis factor RSF1A receptor superfamily, member 1 AIL-6 Human interleukin 6 (interferon, beta 2) IL-8 Human interleukin 8IL-21 Human interleukin 21 IL-22 Human interleukin 22 IL-24 Humaninterleukin 24 IL-26 Human interieukin 26 S100A7 Human S100 calciumbinding protein A7 S100A9 Human S100 calcium binding protein A9 CCL20Human chemokine (C-C motif ligand 20 CXCL1 Human chemokine (C-X-C motif)ligand 1 (melanoma growth stimulating activity, alpha) CXCL5 Humanchemokine (C-X-C motif) ligand 5 LCN2 Human lipocalin 2 DEFB4A Humandefensin, beta 4A

In some embodiments, the one or more target genes comprise C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, the one or more target genes comprise C6orf218.In other cases, the one or more target genes comprise preferentiallyexpressed antigen in melanoma (PRAME).

In some embodiments, one or more target genes from the isolated RNAobtained from a collected skin sample are analyzed, in which the one ormore target genes comprise at least C6orf218, preferentially expressedantigen in melanoma (PRAME), IL-6, IL-8, IL-17A, IL-17C, IL-17F,IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24, IL-26, TNF-α, TNF RSF1A,S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2, DEFB4A, or a combinationthereof. In some instances, from about 1 to about 100, from about 1 toabout 90, from about 1 to about 80, from about 1 to about 70, from about1 to about 60, from about 1 to about 50, from about 1 to about 40, fromabout 1 to about 30, from about 1 to about 20, from about 5 to about100, from about 5 to about 80, from about 5 to about 60, from about 5 toabout 40, from about 5 to about 20, from about 10 to about 100, fromabout 10 to about 80, from about 10 to about 60, from about 10 to about40, from about 20 to about 80, from about 20 to about 60, from about 20to about 40, from about 30 to about 80, from about 30 to about 60, fromabout 40 to about 60, from about 2 to about 10, from about 2 to about 8,or from about 2 to about 6 target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least C6orf218, preferentially expressedantigen in melanoma (PRAME), IL-6, IL-8, IL-17A, IL-17C, IL-17F,IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24, IL-26, TNF-α, TNF RSF1A,S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2, DEFB4A, or a combinationthereof.

In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,20, 25, 30, 40, 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least C6orf218, preferentially expressedantigen in melanoma (PRAME), IL-6, IL-8, IL-17A, IL-17C, IL-17F,IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24, IL-26, TNF-α, TNF RSF1A,S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2, DEFB4A, or a combinationthereof. In some cases, about 1 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), IL-6, IL-8, IL-17A, IL-17C,IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24, IL-26, TNF-α, TNFRSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2, DEFB4A, or acombination thereof. In some cases, about 2 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), IL-6, IL-8,IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A, IL-24,IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5, LCN2,DEFB4A, or a combination thereof. In some cases, about 3 or more targetgenes from the isolated RNA obtained from a collected skin sample areanalyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 4 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 5 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 6 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 7 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 8 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 9 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 10 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 11 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 12 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 13 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 14 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 15 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 20 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 25 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 30 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 40 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof. In some cases, about 50 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218, preferentially expressed antigen in melanoma (PRAME), IL-6,IL-8, IL-17A, IL-17C, IL-17F, IL-17RA, IL-17RC, IL-21, IL-22, IL-23A,IL-24, IL-26, TNF-α, TNF RSF1A, S100A7, S100A9, CCL20, CXCL1, CXCL5,LCN2, DEFB4A, or a combination thereof.

In some embodiments, one or more target genes from the isolated RNAobtained from a collected skin sample are analyzed, in which the one ormore target genes comprise at least one target gene selected fromTable 1. In some instances, from about 1 to about 100, from about 1 toabout 90, from about 1 to about 80, from about 1 to about 70, from about1 to about 60, from about 1 to about 50, from about 1 to about 40, fromabout 1 to about 30, from about 1 to about 20, from about 5 to about100, from about 5 to about 80, from about 5 to about 60, from about 5 toabout 40, from about 5 to about 20, from about 10 to about 100, fromabout 10 to about 80, from about 10 to about 60, from about 10 to about40, from about 20 to about 80, from about 20 to about 60, from about 20to about 40, from about 30 to about 80, from about 30 to about 60, fromabout 40 to about 60, from about 2 to about 10, from about 2 to about 8,or from about 2 to about 6 target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1.

In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,20, 25, 30, 40, 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 1 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 2 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 3 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 4 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 5 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 6 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 7 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 8 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 9 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 10 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 11 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 12 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 13 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 14 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 15 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 20 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 25 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 30 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 40 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1. Insome cases, about 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least one target gene selected from Table 1.

In some embodiments, one or more target genes from the isolated RNAobtained from a collected skin sample are analyzed, in which the one ormore target genes comprise at least C6orf218, preferentially expressedantigen in melanoma (PRAME), or a combination thereof. In someinstances, from about 1 to about 100, from about 1 to about 90, fromabout 1 to about 80, from about 1 to about 70, from about 1 to about 60,from about 1 to about 50, from about 1 to about 40, from about 1 toabout 30, from about 1 to about 20, from about 5 to about 100, fromabout 5 to about 80, from about 5 to about 60, from about 5 to about 40,from about 5 to about 20, from about 10 to about 100, from about 10 toabout 80, from about 10 to about 60, from about 10 to about 40, fromabout 20 to about 80, from about 20 to about 60, from about 20 to about40, from about 30 to about 80, from about 30 to about 60, from about 40to about 60, from about 2 to about 10, from about 2 to about 8, or fromabout 2 to about 6 target genes from the isolated RNA obtained from acollected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof.

In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,20, 25, 30, 40, 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least C6orf218, preferentially expressedantigen in melanoma (PRAME), or a combination thereof. In some cases,about 1 or more target genes from the isolated RNA obtained from acollected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof. In some cases, about 2 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed, in which the one or more target genes comprise atleast C6orf218, preferentially expressed antigen in melanoma (PRAME), ora combination thereof. In some cases, about 3 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, about 4 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), or a combination thereof. In somecases, about 5 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof. In some cases, about 6 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed, in which the one or more target genes comprise atleast C6orf218, preferentially expressed antigen in melanoma (PRAME), ora combination thereof. In some cases, about 7 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, about 8 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), or a combination thereof. In somecases, about 9 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof. In some cases, about 10 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed, in which the one or more target genes comprise atleast C6orf218, preferentially expressed antigen in melanoma (PRAME), ora combination thereof. In some cases, about 11 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, about 12 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), or a combination thereof. In somecases, about 13 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof. In some cases, about 14 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed, in which the one or more target genes comprise atleast C6orf218, preferentially expressed antigen in melanoma (PRAME), ora combination thereof. In some cases, about 15 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, about 20 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), or a combination thereof. In somecases, about 25 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218, preferentially expressed antigen inmelanoma (PRAME), or a combination thereof. In some cases, about 30 ormore target genes from the isolated RNA obtained from a collected skinsample are analyzed, in which the one or more target genes comprise atleast C6orf218, preferentially expressed antigen in melanoma (PRAME), ora combination thereof. In some cases, about 40 or more target genes fromthe isolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218,preferentially expressed antigen in melanoma (PRAME), or a combinationthereof. In some cases, about 50 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218, preferentiallyexpressed antigen in melanoma (PRAME), or a combination thereof.

In some embodiments, one or more target genes from the isolated RNAobtained from a collected skin sample are analyzed, in which the one ormore target genes comprise at least C6orf218. In some instances, fromabout 1 to about 100, from about 1 to about 90, from about 1 to about80, from about 1 to about 70, from about 1 to about 60, from about 1 toabout 50, from about 1 to about 40, from about 1 to about 30, from about1 to about 20, from about 5 to about 100, from about 5 to about 80, fromabout 5 to about 60, from about 5 to about 40, from about 5 to about 20,from about 10 to about 100, from about 10 to about 80, from about 10 toabout 60, from about 10 to about 40, from about 20 to about 80, fromabout 20 to about 60, from about 20 to about 40, from about 30 to about80, from about 30 to about 60, from about 40 to about 60, from about 2to about 10, from about 2 to about 8, or from about 2 to about 6 targetgenes from the isolated RNA obtained from a collected skin sample areanalyzed, in which the one or more target genes comprise at leastC6orf218.

In some cases, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,20, 25, 30, 40, 50 or more target genes from the isolated RNA obtainedfrom a collected skin sample are analyzed, in which the one or moretarget genes comprise at least C6orf218. In some cases, about 1 or moretarget genes from the isolated RNA obtained from a collected skin sampleare analyzed, in which the one or more target genes comprise at leastC6orf218. In some cases, about 2 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218. In some cases, about 3or more target genes from the isolated RNA obtained from a collectedskin sample are analyzed, in which the one or more target genes compriseat least C6orf218. In some cases, about 4 or more target genes from theisolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218. In somecases, about 5 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218. In some cases, about 6 or more targetgenes from the isolated RNA obtained from a collected skin sample areanalyzed, in which the one or more target genes comprise at leastC6orf218. In some cases, about 7 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218. In some cases, about 8or more target genes from the isolated RNA obtained from a collectedskin sample are analyzed, in which the one or more target genes compriseat least C6orf218. In some cases, about 9 or more target genes from theisolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218. In somecases, about 10 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218. In some cases, about 11 or more targetgenes from the isolated RNA obtained from a collected skin sample areanalyzed, in which the one or more target genes comprise at leastC6orf218. In some cases, about 12 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218. In some cases, about 13or more target genes from the isolated RNA obtained from a collectedskin sample are analyzed, in which the one or more target genes compriseat least C6orf218. In some cases, about 14 or more target genes from theisolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218. In somecases, about 15 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218. In some cases, about 20 or more targetgenes from the isolated RNA obtained from a collected skin sample areanalyzed, in which the one or more target genes comprise at leastC6orf218. In some cases, about 25 or more target genes from the isolatedRNA obtained from a collected skin sample are analyzed, in which the oneor more target genes comprise at least C6orf218. In some cases, about 30or more target genes from the isolated RNA obtained from a collectedskin sample are analyzed, in which the one or more target genes compriseat least C6orf218. In some cases, about 40 or more target genes from theisolated RNA obtained from a collected skin sample are analyzed, inwhich the one or more target genes comprise at least C6orf218. In somecases, about 50 or more target genes from the isolated RNA obtained froma collected skin sample are analyzed, in which the one or more targetgenes comprise at least C6orf218.

The subject matter described herein, including the gene expression testsand corresponding transmission of data, in certain aspects, areconfigured to be performed in one or more facilities at one or morelocations. Facility locations are not limited by country and include anycountry or territory. Facility locations are not limited by country andinclude any country or territory. In some instances, one or more stepsof the gene expression test are performed in a different country thananother step of the gene expression test. In some instances, one or moresteps of the gene expression test are performed in a different countrythan one or more steps of the tape stripping aspect. In someembodiments, one or more articles are transferred from one or more ofthe facilities to one or more different facilities for analysis orfurther analysis. An article includes, but is not limited to, one ormore components of the skin sample collection kit, a used adhesive tape,isolated cellular material obtained from a used adhesive tape, processedcellular material, and/or data. Processed cellular material includes,but is not limited to, cDNA reverse transcribed from RNA, amplified RNA,and amplified cDNA. Data includes, but is not limited to, informationregarding the expression level of one or more target genes, informationregarding a gene expression signature, and information regarding adisease, such as melanoma. In some embodiments of the methods, devices,and systems described herein, the analysis is performed and a subsequentdata transmission step will convey or transmit the results of theanalysis. Information regarding a disease, includes, but is not limitedto, identification of a disease state, likelihood of treatment successfor a given disease state, identification of progression of a diseasestate (e.g., invasiveness of melanoma), and identification of a diseasestage (e.g., melanoma stages 0, 1, 2, 3, or 4).

In certain examples, the application of the adhesive tape to a skinsample comprises holding the skin taut and pressing the adhesive tapefirmly on the skin surface while making circular motions on the tape.Between about 1 and about 20, between about 1 and about 15, betweenabout 1 and about 10, between about 1 and about 5, between about 5 andabout 20, between about 10 and about 20, and between about 10 and 15circular motions are made on the tape. In one embodiment, about 15circular motions are made on the tape. In some embodiments, the tape isconfigured to remain on the skin surface for up to 6, 5, 4, 3, 2, and 1minutes. After firm application to the skin, the tape is slowly removedin one direction. In certain aspects, the tape stripping method furthercomprises demarcating the sampled skin region on a second surface of atransparent adhesive tape, wherein the first surface is the skin facingsurface comprising the adhesive matrix. The demarcation indicates thesample region to be processed. The demarcation may be the outline of askin lesion. The marker used for demarcation may be provided in the skinsample collection kit.

In some embodiments of the subject matter described herein, the adhesiveskin sample collection kit comprises a self-addressed package fordelivery of one or more used adhesive tapes to a facility. In someinstances, the package includes a prepaid shipping label. In someembodiments, the facility is a facility which will perform one or morediagnostic steps or procedures involving the cellular material adheredto the one or more used adhesive tapes. In some embodiments, the one ormore diagnostic procedures includes, but is not limited to, any stepperformed in a gene expression test (e.g., a pigmented lesion assay),immunohistochemistry assay, immunophenotyping, ELISA, fluorescent insitu hybridization (FISH), and/or gene sequencing. The facility whereany diagnostic procedure or tape stripping method described herein isperformed is not limited to one country. In some instances, one or morediagnostic procedures or tape stripping methods are performed in one ormore different countries. In some embodiments, a diagnostic procedureincludes data analysis for any step of any diagnostic proceduredescribed herein. In some embodiments, any step of any diagnosticprocedure described herein is performed by a software program or moduleon a computer. In additional or further embodiments, data from any stepof any procedure described herein is transferred to and from facilitieslocated within the same or different countries, including analysisperformed in one facility in a particular location and the data shippedto another location or directly to an individual in the same or adifferent country. In additional or further embodiments, data from anystep of any procedure described herein (including analysis of cellularmaterial such as DNA, RNA, and protein as well as transformed data fromcellular material) is transferred to and/or received from a facilitylocated within the same or different countries, including analysis of adata input, such as cellular material, performed in one facility in aparticular location and corresponding data transmitted to anotherlocation, or directly to an individual, such as data related to thediagnosis, prognosis, responsiveness to therapy, or the like, in thesame or different location or country.

The adhesive skin sample collection kit is configured so that the tapestripping method may be performed by a variety of operators in a varietyof locations. In some embodiments, the method is performed in aclinician's office, an outpatient facility or at a home. The method isnot limited to use in a facility and is configured to be utilized in avariety of locales. The method may is performed by a practitioner, nurseor any individual who has read and understood the instructions for useand is capable of performing the method according to the instructionsfor use sheet.

In some instances, the skin sample collection kit is used in combinationwith skin condition monitoring. For example, images of the skin sampletested are captured and stored on a mobile photoinformatic platform thatmaintains the images with the associated clinical information and datarelating to the skin lesion sampled.

Computer Program

The methods, software, media, and systems disclosed herein comprise atleast one computer processor, or use of the same. The computer processormay comprise a computer program. A computer program may include asequence of instructions, executable in the digital processing device'sCPU, written to perform a specified task. Computer readable instructionsmay be implemented as program modules, such as functions, features,Application Programming Interfaces (APIs), data structures, and thelike, that perform particular tasks or implement particular abstractdata types. In light of the disclosure provided herein, those of skillin the art will recognize that a computer program may be written invarious versions of various languages.

The functionality of the computer readable instructions may be combinedor distributed as desired in various environments. A computer programmay comprise one sequence of instructions. A computer program maycomprise a plurality of sequences of instructions. A computer programmay be provided from one location. A computer program may be providedfrom a plurality of locations. A computer program may include one ormore software modules. A computer program may include, in part or inwhole, one or more web applications, one or more mobile applications,one or more standalone applications, one or more web browser plug-ins,extensions, add-ins, or add-ons, or combinations thereof.

Web Application

A computer program may include a web application. In light of thedisclosure provided herein, those of skill in the art will recognizethat a web application may utilize one or more software frameworks andone or more database systems. A web application may be created upon asoftware framework such as Microsoft® .NET or Ruby on Rails (RoR). A webapplication may utilize one or more database systems including, by wayof non-limiting examples, relational, non-relational, feature oriented,associative, and XML database systems. Suitable relational databasesystems may include, by way of non-limiting examples, Microsoft® SQLServer, mySQL™, and Oracle®. Those of skill in the art will alsorecognize that a web application may be written in one or more versionsof one or more languages. A web application may be written in one ormore markup languages, presentation definition languages, client-sidescripting languages, server-side coding languages, database querylanguages, or combinations thereof. A web application may be written tosome extent in a markup language such as Hypertext Markup Language(HTML), Extensible Hypertext Markup Language (XHTML), or eXtensibleMarkup Language (XML). A web application may be written to some extentin a presentation definition language such as Cascading Style Sheets(CSS). A web application may be written to some extent in a client-sidescripting language such as Asynchronous Javascript and XML (AJAX),Flash® Actionscript, Javascript, or Silverlight®. A web application maybe written to some extent in a server-side coding language such asActive Server Pages (ASP), ColdFusion®, Perl, Java™, JavaServer Pages(JSP), Hypertext Preprocessor (PHP), Python™, Ruby, Tcl, Smalltalk,WebDNA®, or Groovy. A web application may be written to some extent in adatabase query language such as Structured Query Language (SQL). A webapplication may integrate enterprise server products such as IBM® LotusDomino®. A web application may include a media player element. A mediaplayer element may utilize one or more of many suitable multimediatechnologies including, by way of non-limiting examples, Adobe® Flash®HTML 5, Apple® QuickTime®, Microsoft® Silverlight®, Java™, and Unity®.

Mobile Application

A computer program may include a mobile application provided to a mobiledigital processing device. The mobile application may be provided to amobile digital processing device at the time it is manufactured. Themobile application may be provided to a mobile digital processing devicevia the computer network described herein.

A mobile application may be created by techniques known to those ofskill in the art using hardware, languages, and development environmentsknown to the art. Those of skill in the art will recognize that mobileapplications may be written in several languages. Suitable programminglanguages include, by way of non-limiting examples, C, C++, C#,Featureive-C, Java™, Javascript, Pascal, Feature Pascal, Python™, Ruby,VB.NET, WML, and XHTML/HTML with or without CSS, or combinationsthereof.

Suitable mobile application development environments may be availablefrom several sources. Commercially available development environmentsinclude, by way of non-limiting examples, AirplaySDK, alcheMo,Appcelerator®, Celsius, Bedrock, Flash Lite, .NET Compact Framework,Rhomobile, and WorkLight Mobile Platform. Other development environmentsmay be available without cost including, by way of non-limitingexamples, Lazarus, MobiFlex, MoSync, and Phonegap. Also, mobile devicemanufacturers distribute software developer kits including, by way ofnon-limiting examples, iPhone and iPad (iOS) SDK, Android™ SDK,BlackBerry® SDK, BREW SDK, Palm′ OS SDK, Symbian SDK, webOS SDK, andWindows® Mobile SDK.

Those of skill in the art will recognize that several commercial forumsmay be available for distribution of mobile applications including, byway of non-limiting examples, Apple® App Store, Android™ Market,BlackBerry® App World, App Store for Palm devices, App Catalog forwebOS, Windows® Marketplace for Mobile, Ovi Store for Nokia® devices,Samsung® Apps, and Nintendo® DSi Shop.

Standalone Application

A computer program may include a standalone application, which may be aprogram that may be run as an independent computer process, not anadd-on to an existing process, e.g., not a plug-in. Those of skill inthe art will recognize that standalone applications may be oftencompiled. A compiler may be a computer program(s) that transforms sourcecode written in a programming language into binary feature code such asassembly language or machine code. Suitable compiled programminglanguages include, by way of non-limiting examples, C, C++,Featureive-C, COBOL, Delphi, Eiffel, Java™, Lisp, Python™, Visual Basic,and VB .NET, or combinations thereof. Compilation may be oftenperformed, at least in part, to create an executable program. A computerprogram may include one or more executable complied applications.

Web Browser Plug-in

A computer program may include a web browser plug-in. In computing, aplug-in may be one or more software components that add specificfunctionality to a larger software application. Makers of softwareapplications may support plug-ins to enable third-party developers tocreate abilities which extend an application, to support easily addingnew features, and to reduce the size of an application. When supported,plug-ins may enable customizing the functionality of a softwareapplication. For example, plug-ins are commonly used in web browsers toplay video, generate interactivity, scan for viruses, and displayparticular file types. Those of skill in the art will be familiar withseveral web browser plug-ins including, Adobe® Flash® Player, Microsoft®Silverlight®, and Apple® QuickTime®. The toolbar may comprise one ormore web browser extensions, add-ins, or add-ons. The toolbar maycomprise one or more explorer bars, tool bands, or desk bands.

In view of the disclosure provided herein, those of skill in the artwill recognize that several plug-in frameworks may be available thatenable development of plug-ins in various programming languages,including, by way of non-limiting examples, C++, Delphi, Java™ PHP,Python™, and VB .NET, or combinations thereof.

Web browsers (also called Internet browsers) may be softwareapplications, designed for use with network-connected digital processingdevices, for retrieving, presenting, and traversing informationresources on the World Wide Web. Suitable web browsers include, by wayof non-limiting examples, Microsoft® Internet Explorer®, Mozilla®Firefox®, Google® Chrome, Apple® Safari®, Opera Software® Opera®, andKDE Konqueror. The web browser may be a mobile web browser. Mobile webbrowsers (also called mircrobrowsers, mini-browsers, and wirelessbrowsers) may be designed for use on mobile digital processing devicesincluding, by way of non-limiting examples, handheld computers, tabletcomputers, netbook computers, subnotebook computers, smartphones, musicplayers, personal digital assistants (PDAs), and handheld video gamesystems. Suitable mobile web browsers include, by way of non-limitingexamples, Google® Android® browser, RIM BlackBerry® Browser, Apple®Safari®, Palm® Blazer, Palm® WebOS® Browser, Mozilla® Firefox® formobile, Microsoft® Internet Explorer® Mobile, Amazon® Kindle® Basic Web,Nokia® Browser, Opera Software® Opera® Mobile, and Sony′ PSP™ browser.

Software Modules

The medium, method, and system disclosed herein comprise one or moresoftwares, servers, and database modules, or use of the same. In view ofthe disclosure provided herein, software modules may be created bytechniques known to those of skill in the art using machines, software,and languages known to the art. The software modules disclosed hereinmay be implemented in a multitude of ways. A software module maycomprise a file, a section of code, a programming feature, a programmingstructure, or combinations thereof. A software module may comprise aplurality of files, a plurality of sections of code, a plurality ofprogramming features, a plurality of programming structures, orcombinations thereof. The one or more software modules may comprise, byway of non-limiting examples, a web application, a mobile application,and a standalone application. Software modules may be in one computerprogram or application. Software modules may be in more than onecomputer program or application. Software modules may be hosted on onemachine. Software modules may be hosted on more than one machine.Software modules may be hosted on cloud computing platforms. Softwaremodules may be hosted on one or more machines in one location. Softwaremodules may be hosted on one or more machines in more than one location.

Databases

The medium, method, and system disclosed herein comprise one or moredatabases, or use of the same. In view of the disclosure providedherein, those of skill in the art will recognize that many databases maybe suitable for storage and retrieval of geologic profile, operatoractivities, division of interest, and/or contact information of royaltyowners. Suitable databases may include, by way of non-limiting examples,relational databases, non-relational databases, feature orienteddatabases, feature databases, entity-relationship model databases,associative databases, and XML databases. A database may beinternet-based. A database may be web-based. A database may be cloudcomputing-based. A database may be based on one or more local computerstorage devices.

EXAMPLES Example 1: Point of Care Skin Sample Collection

A pigmented lesion located on the hand of a subject is selected for skinsampling. The skin sampling area contains a minimal amount of hair, isnot irritated and has not been previously biopsied. The lesion is about8 mm in size. As exemplified in FIG. 1, the skin sampling area (101)comprising the skin lesion (102) is cleansed with an alcohol pad (103)by a practitioner (104) wearing gloves, and the skin is allowed to airdry for 5 minutes.

A tri-fold skin sample collector is removed from an adhesive skin samplecollection kit exemplified by FIG. 8. FIG. 2 exemplifies the tri-foldskin sample collector (200) comprising a peelable release panel (201)comprising four adhesive tapes (202), a placement area panel (203)comprising a removable liner (204), and a clear panel (205). Thetri-fold skin sample collector has a barcode specific for the subject.The removable liner is removed from the placement area panel (203),exposing four regions (206) designated for the placement of up to fourused adhesive tapes. The four regions of the placement area panel arenot exposed to any skin prior to application of a used tape.

An adhesive tape is removed from the top left side of the peelablerelease panel as exemplified in FIG. 3. The practitioner (104) handlesthe adhesive tape (202) by the tab region (301) so that the matrixmaterial of the central collection area (302) does not come in contactwith a surface prior to skin application. The skin sampling area is heldtaut while the adhesive tape is applied onto the skin sampling area. Anadhesive tape (202) positioned on the cleansed skin sampling area (101)comprising a skin lesion (102) is exemplified in FIG. 4. The adhesivetape is pressed firmly on the skin while making 15 circular motions.FIG. 5 exemplifies the practitioner (104) pressing on the skincomprising a skin lesion (102) while making a circular motion (501). Asexemplified in FIG. 6, the lesion area (102) is demarcated on theadhesive tape (202) using a marker (601) provided in the skin samplecollection kit exemplified in Example 8. The practitioner slowly removesthe used adhesive tape from the skin sampling area by holding the taband pulling in one direction. The used tape (701) comprising a skinsample (702) is placed on the first unoccupied skin collection region(206) of the placement area panel (203) on the tri-fold skin samplecollector (200) as exemplified in FIG. 7. The procedure is repeated withthree additional tapes on the same lesion.

The tri-fold skin sample collector is folded and placed in a packageprovided with the skin sample collection kit. The package containspre-paid postage and is self-addressed to a processing facility.

Example 2: Skin Sample Collection

A pigmented lesion located on the upper back of a subject is selectedfor skin sampling. The skin sampling area contains a minimal amount ofhair, is not irritated and has not been previously biopsied. The lesionis about 15 mm in size. The lesion is sampled utilizing an adhesive skinsample collection kit. The skin sample collection kit includes aninstructions for use sheet (or an instruction manual). The lesion issampled by a capable person who has read and understood the skin samplecollection kit instructions for use sheet.

A pair of gloves is removed from the skin sample collection kit and thefitted onto the person performing the skin sampling procedure. The skinsampling area comprising the pigmented lesion is cleansed with analcohol pad provided in the adhesive skin sample collection kit and theskin is allowed to air dry.

A tri-fold skin sample collector is removed from the adhesive skinsample collection kit. The tri-fold skin sample collector comprises apeelable release panel comprising four adhesive tapes, a placement areapanel comprising a removable liner, and a clear panel. The tri-fold skinsample collector has a barcode specific for the subject. The tri-foldskin sample collector further comprises an area configured for providingpatient information. The tri-fold skin sample collector is labeled withthe subject's name and identifying information. The removable liner isremoved from the placement area panel, exposing four regions designatedfor the placement of up to four used adhesive tapes. The four regions ofthe placement area panel are not exposed to any skin prior toapplication of a used tape.

An adhesive tape is removed from the top left side of the peelablerelease panel. The adhesive tape is handled by the tab region so thatthe matrix material does not come in contact with a surface prior toskin application. The skin is held taut while the adhesive tape isapplied onto the skin sampling area. The adhesive tape is pressed firmlyon the skin while making 10 circular motions. The lesion area isdemarcated on the adhesive tape using a marker provided in the adhesiveskin sample collection kit. The used tape is slowly removed in onedirection by pulling the tab away from the skin. The used tape is placedon the first unoccupied skin collection region of the tri-fold skinsample collector. The skin sample procedure is repeated with threeadditional tapes on the same skin lesion.

The tri-fold skin sample collector comprising 4 used adhesive tapes isfolded and placed in the package provided with the adhesive skin samplecollection kit. The package contains pre-paid postage and isself-addressed to a diagnostics facility.

Example 3: Collection System

The adhesive skin sample collection kit components are stored in acardboard box (800) as exemplified in FIG. 8. The kit contains atri-fold skin sample collector (200) comprising four adhesive tapes,instructions for use sheet, a marking pen, a pre-paid, self-addressedshipping package (801), and a shipping label (802). The tri-fold skinsample collector comprises three panels including a peelable releasepanel comprising the four adhesive tapes, a placement area panelcomprising a removable liner and a clear panel. The tri-fold skin samplecollector further comprises a unique barcode (803) configured toidentify a subject. The adhesive tapes stored on the peelable releasepanel have an expiry date of 2 years from the date of manufacture. Theskin sample collection kit is stored between 10° C. and 30° C. Theinstructions for use sheet (or instruction manual) include allinformation necessary to enable a person to understand and perform themethod. The instructions for use sheet (or instruction manual) includediagrams describing steps of the skin sample collection method.

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes suggested to personsskilled in the art are to be included within the spirit and purview ofthis application and scope of the appended claims.

What is claimed is:
 1. A system for non-invasive collection and analysisof a skin sample, the system comprising: an adhesive skin samplecollection kit comprising at least one adhesive tape, wherein the leastone adhesive tape comprises a collection area comprising an adhesivematrix on a surface, wherein the adhesive matrix comprises a copolymer,and wherein the adhesive matrix is configured to adhere to an effectiveamount of a skin sample for quantifying expression levels of one or moretarget genes in the skin sample.
 2. The system of claim 1, wherein theat least one adhesive tape comprises a tab for handling of the adhesivetape while avoiding handling of the adhesive matrix.
 3. The system ofclaim 1, wherein the polymer comprises a hydrocarbon elastomer.
 4. Thesystem of claim 1, wherein the polymer comprises a styrene copolymercompound.
 5. The system of claim 1, wherein the polymer comprises astyrene-isoprene-styrene (SIS) linear block copolymer compound.
 6. Thesystem of claim 1, wherein the effective amount of the skin samplecomprises from about 1 picogram to about 1 gram of cellular material. 7.The system of claim 1, wherein the effective amount of the skin samplecomprises less than about 500 micrograms.
 8. A method for analyzing askin sample comprising: receiving an adhesive skin sample collection kitcomprising a skin sample adhered to an adhesive matrix, wherein theadhesive skin sample collection kit comprising at least one adhesivetape, wherein the at least one adhesive tape comprises a collection areacomprising the adhesive matrix on a surface, and wherein the adhesivematrix comprises a copolymer; and quantifying expression levels of oneor more target genes in the skin sample.
 9. The method of claim 8,wherein the at least one adhesive tape comprises a tab for handling. 10.The method of claim 9, wherein the tab is sized for avoiding unnecessarycontact with the adhesive matrix of the collection area when handlingthe adhesive tape.
 11. The method of claim 8, wherein the collectionarea is transparent such that a skin sampling area of the subject isvisible after application of the adhesive tape to a skin surface. 12.The method of claim 8, wherein the polymer comprises a hydrocarbonelastomer.
 13. The method of claim 8, wherein the polymer comprises astyrene copolymer compound.
 14. The method of claim 8, wherein thepolymer comprises a styrene-isoprene-styrene (SIS) linear blockcopolymer compound.
 15. The method of claim 8, wherein the adhesive tapedoes not comprise latex, silicone, or both.
 16. The method of claim 8,wherein the adhesive tape is sufficiently flexible to conform to a shapea lesion of a skin surface.
 17. The method of claim 8, wherein theadhesive tape is about 5 to about 100 mm in length.
 18. The method ofclaim 8, wherein the adhesive tape is about 5 to about 40 mm in length.19. The method of claim 8, wherein the adhesive skin sample collectionkit comprises a plurality of adhesive tapes.
 20. The method of claim 8,wherein the skin sample comprises an effective amount of the skinsample, the effective amount of the skin sample comprising from about 1picogram to about 1 gram of cellular material.
 21. The method of claim8, wherein the effective amount of the skin sample comprises less thanabout 500 micrograms of the cellular material.